Plasma anti-Müllerian Hormone Concentration Research Articles

Relationship between plasma anti-Müllerian hormone concentrations in female Holstein calves immediately after birth and milk yield and composition in dams during early to mid-gestation

The energy status of dams during the early stages of gestation may affect ovarian reserve. It was hypothesized that greater milk yield and composition during the early stages of gestation may suppress the number of primordial follicles in the fetuses of dairy cattle. This study aimed to evaluate the anti-Müllerian hormone (AMH), used as an indicator of ovarian reserve, in female Holstein calves immediately after birth. It also examined the relationship between the AMH concentrations and the milk yield and composition of dams during early pregnancy. A total of 85 female Holstein calves (born to first-calving cows [nulliparous heifers], n = 31; second-calving [primiparous] cows, n = 22; and third- or subsequent-calving [multiparous] cows, n = 32) were examined. Blood samples were collected from the female calves after birth and before the first colostrum feeding for the analysis of plasma AMH concentrations. Data on daily milk yield and monthly milk composition during the 1st to 6th month of pregnancy were additionally collected from milking dams. The plasma AMH concentrations ranged from 0.106 to 1542.3 pg/mL, with an average of 291.6 ± 30.5 pg/mL. There was no significant difference observed among the plasma AMH concentrations in female calves born from nulliparous heifers, primiparous cows, multiparous cows. Primiparous cows exhibited numerous negative correlations between the plasma AMH concentrations of their female calves immediately after birth and the daily milk yield and milk composition from the 1st to the 4th month of pregnancy; there was, however, no correlation observed in multiparous cows. Overall, there were large individual variations observed in the plasma AMH concentrations immediately after birth. Both milk yield and the amount of milk components strongly influenced the plasma AMH concentrations in calves immediately after birth in primiparous cows in the growth phase. These factors, conversely, did not affect the plasma AMH concentrations of calves in multiparous cows. The results of this study suggest that high-producing pregnant primiparous cows devote more energy to their growth and lactation, potentially compromising the reproductive potential of their offspring. Thus, improving the feeding management of primiparous cows with the goal of increasing the fertility of their progeny will help high-yielding dairy cows reproduce more successfully.

Abundance of Anti-Muellerian hormone in cat ovaries and correlation of its plasma concentration with animal age, weight and stage of the estrous cycle

In female animals of different species, Anti-Müllerian hormone (AMH) is produced by follicular granulosa cells and has been associated with the ovarian follicle pool. Because concentration of AMH in plasma of ovary-intact female cats is apparently more variable than previously assumed, we have analysed AMH concentration in blood of cats (n = 93) presented for routine ovariectomy and assessed ovarian histology and AMH protein expression in the surgically removed ovaries. We hypothesised that AMH is synthesized only in preantral and small antral follicles and that plasma AMH concentration reflects the antral follicle count (AFC). Corpora lutea were detected in 35% of the female cats, whereas plasma progesterone concentration was ≥1 ng/mL in 57% of the cats. Follicular cysts were present in 15 cats (16%). Positive immunostaining for AMH protein was detected in close to all primordial and antral follicles, ovarian cysts, 70% of corpora lutea and 28% of atretic follicles. Concentration of AMH in plasma averaged 6.8 ± 0.5 ng/mL (range 1.3–21.7 ng/mL). The AFC increased with increasing AMH concentration with a moderate positive correlation between AFC and AMH (r = 0.286, p < 0.01). Plasma AMH concentration was not affected by season or cats’ age, weight, stage of the estrous cycle and presence of follicular cysts. In conclusion, AMH protein is expressed in all endocrine structures of the cat ovary. While AMH is a marker for the presence of ovarian tissue, its usefulness to assess ovarian function in individual female cats is of limited value.

Long-Term Changes in Ovarian Follicles of Gilts Exposed Neonatally to Methoxychlor: Effects on Oocyte-Derived Factors, Anti-Müllerian Hormone, Follicle-Stimulating Hormone, and Cognate Receptors.

In this paper, we investigated the effects of neonatal exposure to methoxychlor (MXC), a synthetic organochlorine used as an insecticide with estrogenic, antiestrogenic, and antiandrogenic activities on ovarian follicles of adult pigs. Piglets were injected with MXC (20 μg/kg body weight) or corn oil (controls) from postnatal Day 1 to Day 10 (n = 5 per group). Then, mRNA expression, protein abundance and immunolocalization of growth and differentiation factor 9 (GDF9), bone morphogenetic protein 15 (BMP15), anti-Müllerian hormone (AMH) and cognate receptors (ACVR1, BMPR1A, BMPR1B, TGFBR1, BMPR2, and AMHR2), as well as FSH receptor (FSHR) were examined in preantral and small antral ovarian follicles of sexually mature gilts. The plasma AMH and FSH levels were also assessed. In preantral follicles, neonatal exposure to MXC increased GDF9, BMPR1B, TGFBR1, and BMPR2 mRNAs, while the levels of AMH and BMP15 mRNAs decreased. In addition, MXC also decreased BMP15 and BMPR1B protein abundance. Regarding small antral follicles, neonatal exposure to MXC upregulated mRNAs for BMPR1B, BMPR2, and AMHR2 and downregulated mRNAs for AMH, BMPR1A, and FSHR. MXC decreased the protein abundance of AMH, and all examined receptors in small antral follicles. GDF9 and BMP15 were immunolocalized in oocytes and granulosa cells of preantral follicles of control and treated ovaries. All analyzed receptors were detected in the oocytes and granulosa cells of preantral follicles, and in the granulosa and theca cells of small antral follicles. The exception, however, was FSHR, which was detected only in the granulosa cells of small antral follicles. In addition, MXC decreased the plasma AMH and FSH concentrations. In conclusion, the present study may indicate long-term effects of neonatal MXC exposure on GDF9, BMP15, AMH, and FSH signaling in ovaries of adult pigs. However, the MXC effects varied at different stages of follicular development. It seems that neonatal MXC exposure may result in accelerated initial recruitment of ovarian follicles and impaired cyclic recruitment of antral follicles.

Assessment of anti-Müllerian hormone in mares’ transitional period and in relation to fertility in elderly mares

Anti-Müllerian Hormone (AMH) has been linked to reproductive tract abnormalities in mares and stallions. This study aimed at evaluating AMH as a biomarker for two reproductive conditions in mares. In the first part of this study, plasma AMH was evaluated as an early indicator of the onset of cyclicity in mares in the transitional period from the anovulatory phase during winter anoestrus to the cyclic phase during the breeding season. Ten mares between 8 and 17 years old were included in the experiment which lasted from mid-February until the end of April. Ovarian activity was monitored with ultrasonography three times per week, the detection of a corpus haemorrhagicum/luteum was documented and antral follicle counts (AFC) were recorded. Blood samples were collected weekly by jugular venipuncture during the whole study period to compare AMH concentrations before and after the first ovulation of the year. The second objective was to evaluate if plasma AMH concentrations in middle-aged mares are linked to fertility and could serve as a prognostic marker in that age group. A total of 41 privately-owned clinically sound mares aged between 12 and 21 years of various breeds were inseminated with fresh or frozen semen. Mares were scanned between day 14 and 20 and the “early pregnancy rate” included only positive pregnancy examinations after the first observed cycle in the season of each mare. Potential associations between the early pregnancy rate in the first cycle and the explanatory factors AMH concentrations, age, status of the mare, stud, development of post-breeding endometritis, number of inseminations and semen type were analysed using logistic regression models. In the first part of the study, correlation between AMH and AFC for the whole study period (P = 0.0002, ρ = 0.55) as well as prior to (P = 0.008, ρ = 0.58) and after the first ovulation (P = 0.0007, ρ = 0.69) were observed. However, AMH concentrations before and after the first ovulation of the year were not statistically different. The second part of the study revealed no association between early pregnancy rate and AMH concentrations or any of the other mentioned factors. In conclusion, this study showed no evidence of a difference between AMH concentrations before and after the first ovulation of the year thus not supporting the use of AMH as a biomarker to predict the onset of cyclicity in mares. We could furthermore not show a relationship between plasma AMH concentrations and early pregnancy rates in this cohort of animals.

Efficacy of a single blood anti-Müllerian hormone (AMH) concentration measurement for the selection of Japanese Black heifer embryo donors in herd breeding programs.

We evaluated the relationship between plasma anti-Müllerian hormone (AMH) concentrations in Japanese Black (JB) heifers at 7−10 months of age and the number of embryos recovered after superovulation treatment in selected ovum pick-up donors, concomitantly with changes in their AMH concentrations before and after parturition. Plasma AMH concentrations in heifers were positively correlated with the total number of follicles (r = 0.647, P < 0.01) and embryos (r = 0.681, P < 0.01) recovered from the animals postpartum, when selected as donor cows, but did not correlate with the total number of transferable embryos. No difference was observed between the plasma AMH concentration at the heifer period and the postpartum period. Additionally, serum AMH concentrations of heifers weakly correlated with the number of follicles and embryos recovered by virgin flush after superovulation treatment at 13−15 months of age. Therefore, a single blood AMH concentration measurement may accelerate intensive JB cattle breeding.

Neonatal Exposure to Agonists and Antagonists of Sex Steroid Receptors Affects AMH and FSH Plasma Level and Their Receptors Expression in the Adult Pig Ovary.

Simple SummaryThe ovarian development and the establishment of ovarian reserve during fetal and/or neonatal life is critical for future reproductive success. Many environmental chemicals are known to negatively affect development and physiology of human and animal ovaries by interfering with endocrine systems, resulting in aberrant reproductive functions. The present study shows the long-term impact of neonatal exposure to agonists and antagonists of sex steroid receptors on AMH and FSH signalling in the ovary of adult pigs. Our findings suggest alteration in ovarian follicle recruitment from ovarian reserve arising from neonatal disruption of androgen/estrogen signalling induced by environmental endocrine active compounds. Everyday use of many endocrine disruptors is already prohibited after their harmful impacts on normal physiology have become known. Nevertheless, market introduction of new chemicals with potential deleterious influence on reproductive physiology has continued. Our outcomes confirm that a neonatal window plays an essential role in the physiological programming of ovarian function in adult pigs. The influence of environmental chemicals on this critical neonatal window needs to be investigated in order to gain a comprehensive view of deleterious interactions between endocrine disrupting chemicals and ovarian function.In this study piglets were injected with testosterone propionate (TP, an androgen), flutamide (FLU, an antiandrogen), 4-tert-octylphenol (OP, an estrogenic compound), ICI 182,780 (ICI, an antiestrogen) or corn oil (controls) between postnatal days 1 and 10 (N = 5/group). Then plasma anti-Müllerian hormone (AMH) and follicle stimulating hormone (FSH) concentration and the expression of their receptors were examined in the adult pig ovary. TP and FLU decreased plasma AMH and FSH concentration. In preantral follicles, TP resulted in upregulation of AMHR2 and FSHR expression, but decreased AMH protein abundance. FLU upregulated AMHR2 expression, while OP increased FSHR mRNA. In small antral follicles, OP upregulated ACVR1 and BMPR1A expression, while FLU increased BMPR1A mRNA. FLU and ICI resulted in upregulation of AMHR2 expression. TP and FLU upregulated AMH expression, while it was downregulated in response to OP or ICI. Moreover, OP and ICI resulted in downregulation of FSHR expression, while FLU decreased FSHR protein abundance. In conclusion, neonatal exposure to either agonist or antagonist of androgen receptor affected AMH and FSH signalling systems in preantral follicles. In small antral follicles these systems were influenced by compounds with estrogenic, antiestrogenic, and antiandrogenic activity. Consequently, these hormonal agents may cause an accelerated recruitment of primordial follicles and affect the cycling recruitment of small antral follicles in pigs.

Anti-Müllerian hormone in grazing dairy cows: Identification of factors affecting plasma concentration, relationship with phenotypic fertility, and genome-wide associations

The objectives of this study were to (1) characterize the distribution and variability of plasma anti-Müllerian hormone (AMH) concentration; (2) evaluate factors associated with phenotypic variation in plasma AMH; (3) examine the associations between categories of plasma AMH and reproductive outcomes [pregnancy to first artificial insemination (P/AI), and pregnancy rates within 21, 42, and 84 d after the mating start date (MSD)]; (4) estimate pedigree and genomic heritability for plasma AMH; and (5) identify and validate SNP associated with phenotypic variation in plasma AMH. Plasma AMH concentration (pg/mL) was determined from a blood sample collected (mean ± standard deviation) 10 ± 2 d after first insemination at detected estrus (IDE) in 2,628 first- and second-parity Irish dairy cows. Overall, plasma AMH had a positively skewed distribution with mean (± standard deviation), median, minimum, and maximum concentrations of 326 ± 231, 268, 15, and 2,863 pg/mL, respectively. Plasma AMH was greatest for Jersey, followed by Holstein × Jersey, Holstein × Norwegian Red, and Holstein cows (410, 332, 284, and 257 pg/mL, respectively). Second-parity cows had greater plasma AMH than first-parity cows (333 vs. 301 pg/mL, respectively). Samples collected at 7 and 8 d after first IDE had lesser plasma AMH than those collected on d 9, 10, 11, 12, and 13 after first IDE (291 and 297 vs. 317, 319, 331, 337, and 320 pg/mL). Plasma AMH was not associated with either body condition score at first IDE or the interval from calving to MSD. Cows were categorized into low (≤150 pg/mL; n = 526; lowest 20%), intermediate (>150 to ≤461 pg/mL; n = 1,576; intermediate 60%), and high AMH (>461 pg/mL; n = 526; highest 20%) groups based on plasma AMH, and associations with reproductive outcomes were tested. Cows with high and intermediate plasma AMH had 1.42- and 1.51-times-greater odds of becoming pregnant within 84 d after the MSD than those with low plasma AMH (90.3 and 90.8 vs. 86.8%, respectively); however, P/AI and pregnancy rate within 21 and 42 d after the MSD did not differ among AMH categories. Plasma AMH was moderately heritable (pedigree heritability of 0.40 ± 0.06 and genomic heritability of 0.45 ± 0.05), and 68 SNP across Bos taurus autosomes 7 and 11 were associated with phenotypic variation in plasma AMH. Out of 68 SNP, 42 were located in a single quantitative trait locus on Bos taurus autosome 11 that harbored 6 previously identified candidate genes (NR5A1, HSPA5, CRB2, DENND1A, NDUFA8, and PTGS) linked to fertility-related phenotypes in dairy cows.

Association of Anti-Müllerian Hormone concentrations between the pregnancy rates and pregnancy continuity of cows in different age groups

The potential relationships between Anti-Müllerian Hormone (AMH) concentrations and fertility were investigated by examining pregnancy rates and early pregnancy loss in different age groups of cows. Holstein heifers (17.35 ± 1.35 months, n = 20), young cows (3.25 ± 1.02 years, n = 20), and old cows (6.7 ± 0.80 years, n = 20) were synchronised and time fixed inseminated. A single blood sample per animal was taken during oestrus just before artificial insemination (AI), (Day 0) for the AMH analysis. The highest plasma AMH concentrations were determined in the heifer group (149.01 ± 12.62 pg/ml, P &amp;lt; 0.001) in the study. The AMH concentrations and conception rates decreased with age. The Day-0-AMH concentrations were higher in the pregnant animals at Day 20 after AI than in the non-pregnant animals (P &amp;lt; 0.05) in each group. The mean Day-0-AMH concentrations in the pregnant animals at Day 60 after AI was higher (P &amp;lt; 0.001) than the AMH concentrations in those with a detected pregnancy loss between D&lt;sub&gt;20&lt;/sub&gt; and D&lt;sub&gt;60&lt;/sub&gt;, and also in those with a pregnancy loss and not being pregnant by Day 20 considered together. When each group was assessed on its own, based on the pregnancy results of Day 60, the Day-0-AMH concentrations were found to be significantly higher in the pregnant heifers than in the non-pregnant heifers and heifers with a pregnancy loss (P &amp;lt; 0.05). In conclusion, the AMH analysis might provide valuable information on the reproductive efficiency of the animals in a herd.

A bovine-specific FSH enzyme immunoassay and its application to study the role of FSH in ovarian follicle development during the postnatal period

The primary aim of this study was to develop a FSH enzyme immunoassay (EIA) for the bovine species. The newly developed EIA was validated for FSH determination in bovine plasma by comparison with an existing bovine FSH radioimmunoassay. The EIA detected bovine FSH with a high sensitivity (0.1 ng/ml). Cross-reactivity of the EIA was 0.01% with bovine LH, 51% with ovine FSH, <0.1% with porcine FSH and <0.01% with equine FSH. Using this EIA on different time series of plasma in cows, we have confirmed the presence of a FSH pre-ovulatory peak at estrus, of periodic FSH fluctuations accompanying the waves of terminal follicular development, and of FSH pulses, mainly asynchronous with LH ones, in the peri-ovulatory phase of the cycle. In a second objective, the EIA was used to assess the role of FSH in regulating the development of ovarian follicles up to the small antral stage in young calves. To answer this question, six calves were submitted to weekly blood sampling during their first 3 months of life, and FSH changes were studied concomitantly to those of anti-Müllerian hormone (AMH), a well-established endocrine marker of the ovarian population of small antral follicles in cows. In the ovaries of 3-month calves, the population of 3 to 5 mm follicles contained the highest intra-follicular AMH amounts, and the number of 3 to 5 mm follicles on ovaries was closely correlated with AMH concentrations in the plasma of calves at this age (rs = 0.94). Before 3 months of age, only two out of six calves showed a clear postnatal FSH peak in plasma, and no correlation was found between plasma FSH and AMH concentrations. These results indicate that female calves undergo different patterns of FSH secretion and that postnatal activation of follicular growth up to the small antral stage appears independent and not directly related to circulating FSH levels.

Effects of genetic background on responses to superovulation in Japanese Black cattle.

We investigated the effects of genetic background on the responses to superovulation in Japanese Black cattle. The genotype frequencies of GRIA1 and FSHR relating to ovulation and follicular development in each of the major bloodlines—Tajiri, Fujiyoshi, and Kedaka—were analyzed. The Tajiri line had the lowest frequency of G allele homozygosity of c.710A>G in GRIA1 among the three bloodlines, and deviation from Hardy–Weinberg equilibrium was detected. Genotype frequencies of c.337C>G, c.871A>G, and c.1973C>G in FSHR were in Hardy–Weinberg equilibrium in all bloodlines. The results of generalized linear mixed-model analyses showed that farm, levels of plasma anti-Müllerian hormone (AMH) concentration, age in months, repeated superovulation, c.337C>G in FSHR, and bloodlines had significant effects on the responses to superovulation. The number of transferable embryos in the group heterozygous for c.337C>G in FSHR was significantly higher than that in the group homozygous for the C allele. The Kedaka line showed a significantly higher number of ova/embryos, fertilized embryos, and transferable embryos than the Tajiri and Fujiyoshi lines. The concentration of circulating AMH is a useful endocrine marker for antral follicle counts. This study revealed the effects of genetic background on the responses to superovulation using levels of plasma AMH concentration as a covariate. The prominent effect of genetic background on superovulation in the Kedaka line requires additional studies to confirm the genomic regions and polymorphisms that are involved in the trait.

Plasma anti-Müllerian hormone profile in heifers from birth through puberty and relationship with puberty onset.

The current study aimed to define the plasma profile of anti-Müllerian hormone (AMH) and follicle stimulating hormone (FSH) in heifers during postnatal life until achieving puberty, as defined by plasma progesterone (P4) profile, to demonstrate a relationship between AMH and age of puberty onset. Blood samples collected from 11 Japanese Black female calves within 1 week after birth (W0) and then biweekly until the sixth week after puberty (WP 6) were assayed for AMH, FSH, and P4. The heifers were classified into two groups based on age of puberty onset: ≤42 weeks (early puberty group; EP, n =4) and ≥44 weeks (late puberty group; LP, n =7). Minimal plasma AMH concentration occurring at W 0 gradually increased to its peak level by W 10 (fourfold higher than W 0; P<0.01) before gradually declining to a steady plateau 6 weeks before puberty (WP -6). The AMH peak was preceded by a significant rise in plasma FSH at W 4, W 6, and W 8 compared with W0. Plasma AMH at W 16 positively correlated with WP 4 and WP 6 (r =0.69 and 0.71, respectively; P<0.05). Overall plasma AMH and FSH was significantly higher and lower in EP compared with LP, respectively. In conclusion, heifers exhibit a characteristic plasma AMH profile during postnatal life, such that plasma AMH at an early prepubertal age could be a biomarker for precocious puberty and postpubertal AMH levels.

Relationship between plasma anti-Müllerian hormone concentrations during the rearing period and subsequent embryo productivity in Japanese black cattle

To use plasma anti-Müllerian hormone (AMH) concentrations as early-stage markers for donor cow selection, we investigated the relationship between plasma AMH concentrations in Japanese black heifers and subsequent embryo productivity following superovulation treatment. Plasma AMH and nonesterified fatty acid (NEFA) concentrations in 6 heifers were evaluated once per month from 3 mo before successful AI for primiparity to 3 mo postpartum. Following calving, embryo collection by superovulation treatment was performed at 3- to 4-mo intervals. There were no significant differences in plasma AMH concentrations between the time points throughout the study period. There were, however, significant inter-animal differences in plasma AMH concentrations (P < 0.05). These findings suggest that plasma AMH concentrations were stable over time and individually specific. There were significant positive correlations between plasma AMH concentrations before AI and embryo productivity variables, including the number of ova/embryos (number of transferable embryos, degenerated embryos, and unfertilized oocytes) and numbers/proportions of fertilized and transferable embryos. There was no significant correlation between plasma AMH and NEFA concentrations throughout the study period. These findings reveal that plasma AMH concentrations during the rearing period can be used to predict subsequent embryo productivity following superovulation treatment, suggesting that these concentrations are useful early-stage markers for selecting donor cows.

Positive correlations of age and parity with plasma anti-Müllerian hormone concentrations in Japanese Black cows.

Anti-Müllerian hormone (AMH) is secreted from the preantral and small antral follicles. It regulates follicle development and inhibits follicular atresia. This study examined how age, parity, and time after parturition affect plasma AMH concentrations in Japanese Black cows. We measured plasma AMH concentrations in primiparous, secundiparous, and multiparous (third parity or higher) cows at four time points: day 2 (day 0 = parturition), day 8, 2 days before first postpartum ovulation (pre-1stOv), and 12 days after first ovulation (post-1stOV). We observed a positive correlation between plasma AMH concentration and age (in months) and parity on day 2, day 8, and post-1stOV, but not on pre-1stOv. The multiparous cows had higher AMH concentrations than primiparous cows throughout the postpartum period (P < 0.05). Our results indicate that age and parity significantly influence plasma AMH concentrations in Japanese Black cows during the voluntary waiting period.

Anti-Müllerian hormone (AMH) as a predictor of antral follicle population in heifers.

Antral follicular population (AFP) and anti-Müllerian hormone (AMH) concentration are strongly associated. Thus, analyzing AMH concentration is a reliable method to predict animals with high AFP, an important feature to select donor cows for embryos and oocytes. However, not much is known about this parameter in heifers, particularly in crossbred animals. The aim of this study was to assess AFP in heifers and cows and its relation to serum AMH levels. Bos taurus indicus cows (36–60-months-old; n = 30) and heifers (12–14-months-old; n = 17) of the same breed were evaluated. A single procedure of follicular counting was performed by ultrasound for each animal. Random sampling was performed to analyze serum AMH concentration. Cows showed higher AFP than heifers; nonetheless, plasma AMH concentration did not differ between the categories. A high correlation of AFP with plasma AMH concentration was observed in Girolando heifers. Thus, it is suggested that AMH concentration could be a reliable and less invasive method for selecting heifers with high AFP.

Long-term changes in plasma anti-Müllerian hormone concentration and the relationship with superovulatory response in Japanese Black cattle.

The concentration of circulating anti-Müllerian hormone (AMH) in cattle is a useful endocrine marker for ovarian response to superovulation. Although the AMH concentration undergoes little variation throughout the estrous cycle, its long-term changes remain incompletely understood. Here, we investigated the relationship between superovulation response and plasma AMH concentration in Japanese Black cattle and the long-term changes in plasma AMH concentration of embryo donor cows and heifers. The median, 25th percentile, and 75th percentile of AMH concentrations in 222 mature animals were 0.265, 0.118, and 0.488 ng/ml, respectively. The numbers of ova/embryos, fertilized embryos, and transferable embryos in a total of 295 superovulations were significantly different among the H (AMH ≥ 0.488 ng/ml), M (AMH 0.487–0.119 ng/ml), and L (AMH ≤ 0.118 ng/ml) groups. AMH concentrations during repeated superovulation in ten donor cows were significantly decreased after the third treatment. In heifers, the highest AMH concentration was observed in individuals during 2–13 months of age, with considerable individual variability. AMH concentrations of heifers at 10 or 11 months correlated with the number of ova/embryos during superovulation at 13–18 months (r = 0.641, P < 0.05). These results suggest that the 25th and 75th percentile values of AMH concentration would give a useful rough estimate of ovarian response; however, repeated superovulation may reduce the predictive accuracy of single measurements of AMH concentration. It would be possible to evaluate AMH concentration in heifers after approximately 11 months of age.

Clinical diagnosis of bovine granulosa cell tumour in a Holstein cow using plasma anti-Müllerian hormone concentration: a case report

ABSTRACTA 23-month-old Holstein cow had a multi-cystic and enlarged left ovary detected by rectal palpation and ultrasonography. The plasma anti-Müllerian hormone (AMH) concentration was 36.5 ng/ml, which is higher than normal. From the clinical history and plasma AMH levels, a granulosa cell tumour (GCT) was diagnosed. Affected ovary was removed by ovariectomy, and GCT was confirmed with histopathological findings. The cow was artificially inseminated at 145 days after ovariectomy, and conceived. The plasma AMH concentration declined at 224 days after removal of affected ovary to less than 0.1 ng/ml, and recovery of fertility was achieved. In conclusion, measurement of plasma AMH levels combined with rectal palpation and ultrasonography examinations could be useful in clinical diagnosis of bovine GCT.

Changes in peripheral anti-Müllerian hormone concentration and their relationship with testicular structure in beef bull calves

The aim of this study was to clarify the time-course of changes in anti-Müllerian hormone (AMH) and testosterone (T) concentrations in peripheral blood and to determine the relationships between blood AMH concentration and testicular development during the early postnatal and prepubertal periods in beef bull calves. A total of 17 Japanese Black bull calves were enrolled in this study. The wk in which the calf was born (within 6 d after birth) was defined as M 0. Blood samples were taken once in every mo from M 0 to M 6 from each bull calf, and plasma AMH and T concentrations were determined. Of the 17 calves, 10 were castrated at 6 mo of age (prepuberty) and the right testis was histologically examined. Plasma AMH concentration (means ± SE) at M 0, 1, and 2 were 123.5 ± 9.8, 189.6 ± 18.7, and 254.6 ± 14.1 ng/mL, respectively. From M 0 through M 2, plasma AMH concentration was significantly greater each mo than in the previous mo (P < 0.05); however, plasma AMH concentration significantly decreased over the last 3 mo of the study (P < 0.05). The average age at which plasma AMH concentration was the highest was 2.3 ± 0.1 mo of age. Plasma T concentration significantly increased from M 0 (0.18 ± 0.02 ng/mL) until M 6 (6.52 ± 1.41 ng/mL). Plasma AMH and T concentrations at M 4, 5, and 6 were significantly negatively correlated (P < 0.05). Linear regression did not reveal a significant relationship between Sertoli or Leydig cell numbers and plasma AMH or T concentrations, respectively. In conclusion, blood AMH concentration peaks at 2 mo of age and is negatively correlated with blood T concentration from 4 to 6 mo of age. Although prepubertal blood AMH or T concentrations did not reflect Sertoli or Leydig cell numbers at the end of the prepubertal period, blood AMH concentration may be indicative of abnormal Sertoli cells function.

The use of anti-Müllerian hormone as diagnostic for gonadectomy status in dogs

In the veterinary practice, there is a need for a diagnostic tool to check the gonadal status in female dogs because it may be difficult to determine whether a female animal has been spayed or whether there are ovarian remnants. Although less prevalent, a similar situation pertains to male dogs. Anti-Müllerian hormone (AMH) is an important regulator of gonadal function and is a specific gonadal product that can be determined in circulation. The objective of this study was to develop and test a canine blood AMH assay as a diagnostic tool to determine the presence of functional gonadal tissue in dogs. A prospective study with a training-validation set paradigm was used. A canine AMH assay was developed and serum and plasma AMH concentrations were determined in blood samples from 46 intact female dogs, 48 spayed females, 50 intact males, and 48 castrated males collected at two separate institutes. Using a training-validation set paradigm, it was found that using cutoff values of 1.1 ng/mL (female) and 5.5 ng/mL (male) AMH, the assay reported excellent specificity and sensitivity of 100% and 90% in female dogs, and good specificity and sensitivity of 100% and 76%, in male dogs, respectively. The sensitivity in male dogs could be further enhanced by including a serum testosterone determination. This newly developed canine AMH assay is a valuable diagnostic tool to determine gonadal status in veterinary medicine.

Anti-Müllerian hormone serum concentrations of women with germline BRCA1 or BRCA2 mutations.

STUDY QUESTIONDo women with BRCA1 or BRCA2 mutations have reduced ovarian reserve, as measured by circulating anti-Müllerian hormone (AMH) concentration?SUMMARY ANSWERWomen with a germline mutation in BRCA1 have reduced ovarian reserve as measured by AMH.WHAT IS KNOWN ALREADYThe DNA repair enzymes encoded by BRCA1 and BRCA2 are implicated in reproductive aging. Circulating AMH is a biomarker of ovarian reserve and hence reproductive lifespan.STUDY DESIGN, SIZE, DURATIONThis was a cross-sectional study of AMH concentrations of 693 women at the time of enrolment into the Kathleen Cuningham Foundation Consortium for research in the Familial Breast Cancer (kConFab) cohort study (recruitment from 19 August 1997 until 18 September 2012). AMH was measured on stored plasma samples between November 2014 and January 2015 using an electrochemiluminescence immunoassay platform.PARTICIPANTS/MATERIALS, SETTING, METHODSEligible women were from families segregating BRCA1 or BRCA2 mutations and had known mutation status. Participants were aged 25–45 years, had no personal history of cancer, retained both ovaries and were not pregnant or breastfeeding at the time of plasma storage. Circulating AMH was measured for 172 carriers and 216 non-carriers from families carrying BRCA1 mutations, and 147 carriers and 158 non-carriers from families carrying BRCA2 mutations. Associations between plasma AMH concentration and carrier status were tested by linear regression, adjusted for age at plasma storage, oral contraceptive use, body mass index and cigarette smoking.MAIN RESULTS AND THE ROLE OF CHANCEMean AMH concentration was negatively associated with age (P < 0.001). Mutation carriers were younger at blood draw than non-carriers (P ≤ 0.031). BRCA1 mutation carriers had, on average, 25% (95% CI: 5%–41%, P = 0.02) lower AMH concentrations than non-carriers and were more likely to have AMH concentrations in the lowest quartile for age (OR 1.84, 95% CI: 1.11–303, P = 0.02). There was no evidence of an association between AMH concentration and BRCA2 mutation status (P = 0.94).LIMITATIONS, REASONS FOR CAUTIONAMH does not directly measure the primordial follicle pool. The clinical implications of the lower AMH concentrations seen in BRCA1 mutation carriers cannot be assessed by this study design.WIDER IMPLICATIONS OF THE FINDINGSWomen with a germline mutation in BRCA1 may have reduced ovarian reserve. This is consistent with other smaller studies in the literature and has potential implications for fertility and reproductive lifespan.STUDY FUNDING/COMPETING INTEREST(S)kConFab is supported by a grant from the Australian National Breast Cancer Foundation, and previously by the National Health and Medical Research Council (NHMRC), the Queensland Cancer Fund, the Cancer Councils of New South Wales, Victoria, Tasmania and South Australia, and the Cancer Foundation of Western Australia. K.A.P. is an Australian National Breast Cancer Foundation Practitioner Fellow. J.L.H. is a NHMRC Senior Principal Research Fellow. M.H. is a NHMRC Practitioner Fellow. R.A.A. reports personal fees from Roche Diagnostics & Beckman Coulter outside the submitted work and C.S. reports other earnings from Melbourne IVF outside the submitted work. The remaining authors have nothing to declare and no conflicts of interest.

Molecular changes in the equine follicle in relation to variations in antral follicle count and anti-Müllerian hormone concentrations.

The wide variation in circulating anti-Müllerian hormone (AMH) concentrations between mares is attributed to differences in antral follicle count (AFC) which may reflect follicular function. There are few data regarding variations in AFC and associated regulatory factors for AMH in the equine follicle during follicular development. To examine molecular and hormonal differences in the equine follicle in relation to variations in AFC and circulating AMH concentrations during follicular development and to identify genes co-expressed with AMH in the equine follicle. Observational study. Plasma AMH concentrations and AFC were determined in 30 cyclic mares. Granulosa cells, theca cells and follicular fluid were recovered from growing (n = 17) or dominant follicles (n = 13). The expression of several genes, known to be involved in folliculogenesis and steroidogenesis, was examined using real-time reverse transcriptase polymerase chain reaction and immunohistochemistry. Intrafollicular oestradiol and AMH concentrations were determined by immunoassay. Within growing follicles, the expression of AMH, AMHR2, ESR2 and INHA in granulosa cells was positively correlated with AFC and plasma AMH concentrations. In addition, the expression of ESR1 and FSHR was positively associated with plasma AMH concentrations. No significant associations were detected in dominant follicles. Furthermore, there was no association between AMH or oestradiol concentrations in follicular fluid and variations in AFC. Finally, the expression of AMH and genes co-expressed with AMH (AMHR2, ESR2 and FSHR) in granulosa cells as well as intrafollicular AMH concentrations decreased during follicular development while intrafollicular oestradiol concentrations increased and were inversely related to intrafollicular AMH concentrations. This study indicates that variations in AFC and circulating AMH concentrations are associated with molecular changes in the growing equine follicle.